Use of hydroxysafflor yellow A in treating diseases related to platelets

Abstract

This invention relates to the use of hydroxysaffron yellow A in the treatment of platelet-related diseases. Specifically, this invention provides a use of hydroxysaffron yellow A for preparing compositions for the prevention and / or treatment of platelet-related diseases. This invention discovers that hydroxysaffron yellow A can reduce platelet count and the ratio of large platelets, as well as inhibit platelet aggregation and agglutination, for the prevention and treatment of diseases associated with elevated platelet counts such as thrombocytosis, diseases associated with elevated large platelet ratios, and diseases associated with platelet activation, thereby preventing and treating platelet-related diseases.

Description

Technical Field

[0001] This invention relates to the field of pharmaceuticals, and more specifically to the use of hydroxysaffron yellow A in the treatment of platelet-related diseases. Background Technology

[0002] Platelets are a major component of the circulatory system and are key cells for normal physiological hemostasis. However, under the stimulation of certain factors such as drugs or pathological cytokines, platelets are prone to pathological changes such as increased number, elevated large platelet ratio, or excessive platelet activation, leading to diseases such as thrombocytosis, elevated large platelet ratio, and pathological thrombosis. This results in nonspecific symptoms such as fatigue and weakness, and an increased incidence of thrombosis, such as arterial and venous thrombosis, which can cause adverse symptoms such as headache, visual symptoms, and erythromelalgia. If treatment is inappropriate or delayed, it can further induce diseases such as acute myocardial infarction, cerebral infarction, and coronary atherosclerosis, thus endangering the patient's health. Pathological changes in platelets play a crucial role in the development of diseases such as thrombocytosis, elevated large platelet ratio, and excessive platelet activation. How to prevent and treat diseases such as thrombocytosis, elevated large platelet ratio, and excessive platelet activation caused by pathological changes in platelets remains a hot research topic.

[0003] Therefore, there is a need in this field to develop a drug that can effectively treat diseases such as thrombocytosis, elevated large platelet ratio, and excessive platelet activation caused by pathological changes in platelets. Summary of the Invention

[0004] The purpose of this invention is to provide the use of hydroxysaffron yellow pigment A in the prevention and treatment of platelet-related diseases.

[0005] The first aspect of the present invention provides the use of hydroxysaffron yellow pigment A for preparing compositions for the prevention and / or treatment of platelet-related diseases.

[0006] Preferably, the prevention and / or treatment of platelet-related diseases includes one or more selected from the group consisting of: (i) prevention and / or treatment of thrombocytosis; (ii) reduction of platelet count; (iii) prevention and / or treatment of diseases associated with elevated platelet count; (iv) inhibition of platelet activation; (v) prevention and / or treatment of diseases associated with platelet activation; (vi) inhibition of platelet aggregation and / or clustering; (vii) prevention and / or treatment of diseases associated with platelet aggregation and / or clustering; (viii) reduction of large platelet ratio; (ix) prevention and / or treatment of elevated large platelet ratio; and / or (x) prevention and / or treatment of diseases associated with elevated large platelet ratio.

[0007] Preferably, the composition is a pharmaceutical composition.

[0008] Preferably, the composition further includes a pharmaceutically acceptable carrier.

[0009] Preferably, the dosage form of the composition is a solid dosage form, a semi-solid dosage form, or a liquid dosage form.

[0010] Preferably, the dosage form of the composition is an oral preparation, a topical preparation, or an injectable preparation.

[0011] Preferably, the dosage form of the composition is an emulsion, tablet, capsule, oral liquid, granule, powder, syrup, or injection.

[0012] Preferably, the injectable preparation is an intravenous injection preparation, an intramuscular injection preparation, a subcutaneous injection preparation, an intradermal injection preparation, or an intraperitoneal injection preparation.

[0013] Preferably, the composition is administered to humans or non-human mammals.

[0014] Preferably, the platelets are human or non-human mammal platelets.

[0015] Preferably, the platelets include platelets in blood.

[0016] Preferably, the object of the prevention and / or treatment is a human or non-human mammal.

[0017] Preferably, the subject is a subject that has already been given adrenaline.

[0018] Preferably, the composition is administered to a subject who has already received adrenaline.

[0019] Preferably, the subject of the prevention and / or treatment is a subject who has already received adrenaline.

[0020] Preferably, the object is a human or a non-human mammal.

[0021] Preferably, the non-human mammal is a mouse, rat, rabbit, cow, sheep, dog, or horse.

[0022] Preferably, the prevention and / or treatment of thrombocytosis includes preventing and / or treating thrombocytosis by reducing the number of platelets.

[0023] Preferably, the prevention and / or treatment of diseases associated with elevated platelet counts includes preventing and / or treating diseases associated with elevated platelet counts by reducing platelet counts.

[0024] Preferably, the prevention and / or treatment of platelet activation-related diseases includes preventing and / or treating platelet activation-related diseases by inhibiting platelet aggregation and / or aggregation.

[0025] Preferably, the prevention and / or treatment of elevated platelet ratio includes preventing and / or treating elevated platelet ratio by reducing the platelet ratio.

[0026] Preferably, the thrombocytosis includes thrombocytosis induced by adrenaline administration.

[0027] Preferably, the reduction in platelet count includes reducing the platelet count in the subject of adrenaline administration.

[0028] Preferably, the platelet count of the subject receiving the adrenaline is elevated.

[0029] Preferably, the increase in platelet count includes an increase in platelet count induced by adrenaline administration.

[0030] Preferably, the platelet activation includes platelet activation induced by adrenaline administration.

[0031] Preferably, the platelet aggregation and / or aggregation includes platelet aggregation and / or aggregation induced by adrenaline administration.

[0032] Preferably, the reduction in large platelet ratio includes reducing the large platelet ratio in the subject of adrenaline administration.

[0033] Preferably, the large platelet ratio is increased in the recipient of the adrenaline.

[0034] Preferably, the increased platelet-to-large platelet ratio includes an increased platelet-to-large platelet ratio induced by adrenaline administration.

[0035] Preferably, the administration is oral or injectable.

[0036] Preferably, the injection administration is intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection, or intraperitoneal injection.

[0037] Preferably, the disease includes thrombosis.

[0038] Preferably, the composition further includes anagrelide or a pharmaceutically acceptable salt thereof.

[0039] Preferably, the pharmaceutically acceptable salt is a hydrochloride salt.

[0040] Preferably, the anagrelide or a pharmaceutically acceptable salt thereof comprises anagrelide hydrochloride.

[0041] Preferably, the weight ratio of the hydroxysaffron yellow A to the anagre or its pharmaceutically acceptable salt is 0.1-40:1, more preferably 0.3-30:1, more preferably 0.5-20:1, more preferably 0.5-15:1, more preferably 1-10:1, more preferably 1-8:1, more preferably 1-5:1, more preferably 2-4:1, more preferably 2.5-3.5:1, and more preferably 3:1.

[0042] A second aspect of the invention provides a composition comprising hydroxysafflower yellow A and anagrelide or a pharmaceutically acceptable salt thereof.

[0043] Preferably, the composition is a pharmaceutical composition.

[0044] Preferably, the composition further includes a pharmaceutically acceptable carrier.

[0045] Preferably, the dosage form of the composition is a solid dosage form, a semi-solid dosage form, or a liquid dosage form.

[0046] Preferably, the dosage form of the composition is an oral preparation, a topical preparation, or an injectable preparation.

[0047] Preferably, the dosage form of the composition is an emulsion, tablet, capsule, oral liquid, granule, powder, syrup, or injection.

[0048] Preferably, the injectable preparation is an intravenous injection preparation, an intramuscular injection preparation, a subcutaneous injection preparation, an intradermal injection preparation, or an intraperitoneal injection preparation.

[0049] Preferably, the pharmaceutically acceptable salt is a hydrochloride salt.

[0050] Preferably, the anagrelide or a pharmaceutically acceptable salt thereof comprises anagrelide hydrochloride.

[0051] Preferably, the weight ratio of the hydroxysaffron yellow A to the anagre or its pharmaceutically acceptable salt is 0.1-40:1, more preferably 0.3-30:1, more preferably 0.5-20:1, more preferably 0.5-15:1, more preferably 1-10:1, more preferably 1-8:1, more preferably 1-5:1, more preferably 2-4:1, more preferably 2.5-3.5:1, and more preferably 3:1.

[0052] A third aspect of the present invention provides a medicine box, the medicine box comprising:

[0053] (a) A first formulation, wherein the first formulation comprises hydroxysaffron yellow A;

[0054] (b) A second formulation comprising anagrelide or a pharmaceutically acceptable salt thereof.

[0055] Preferably, the preparation is a pharmaceutical preparation.

[0056] Preferably, the formulation further includes a pharmaceutically acceptable carrier.

[0057] Preferably, the first and second formulations are either independent formulations or combined formulations.

[0058] Preferably, the dosage form of the first formulation and / or the second formulation is a solid dosage form, a semi-solid dosage form, or a liquid dosage form.

[0059] Preferably, the dosage form of the first formulation and / or the second formulation is an oral formulation, a topical formulation, or an injectable formulation.

[0060] Preferably, the dosage form of the composition is an emulsion, tablet, capsule, oral liquid, granule, powder, syrup, or injection.

[0061] Preferably, the injectable preparation is an intravenous injection preparation, an intramuscular injection preparation, a subcutaneous injection preparation, an intradermal injection preparation, or an intraperitoneal injection preparation.

[0062] Preferably, the pharmaceutically acceptable salt is a hydrochloride salt.

[0063] Preferably, the anagrelide or a pharmaceutically acceptable salt thereof comprises anagrelide hydrochloride.

[0064] Preferably, the weight ratio of the hydroxysaffron yellow A to the anagre or its pharmaceutically acceptable salt is 0.1-40:1, more preferably 0.3-30:1, more preferably 0.5-20:1, more preferably 0.5-15:1, more preferably 1-10:1, more preferably 1-8:1, more preferably 1-5:1, more preferably 2-4:1, more preferably 2.5-3.5:1, and more preferably 3:1.

[0065] Preferably, the medicine box also includes an instruction manual, which describes the combined administration of the first and second formulations.

[0066] The fourth aspect of the present invention provides the use of a composition as described in the second aspect of the present invention or a medicine box as described in the third aspect of the present invention for preparing a formulation or medicine for the prevention and / or treatment of platelet-related diseases.

[0067] Preferably, the prevention and / or treatment of platelet-related diseases includes one or more selected from the group consisting of: (i) prevention and / or treatment of thrombocytosis; (ii) reduction of platelet count; (iii) prevention and / or treatment of diseases associated with elevated platelet count; (iv) inhibition of platelet activation; (v) prevention and / or treatment of diseases associated with platelet activation; (vi) inhibition of platelet aggregation and / or clustering; (vii) prevention and / or treatment of diseases associated with platelet aggregation and / or clustering; (viii) reduction of large platelet ratio; (ix) prevention and / or treatment of elevated large platelet ratio; and / or (x) prevention and / or treatment of diseases associated with elevated large platelet ratio.

[0068] Preferably, the preparation is a pharmaceutical preparation.

[0069] Preferably, the formulation further includes a pharmaceutically acceptable carrier.

[0070] Preferably, the dosage form of the preparation is a solid dosage form, a semi-solid dosage form, or a liquid dosage form.

[0071] Preferably, the dosage form of the preparation is an oral preparation, a topical preparation, or an injectable preparation.

[0072] Preferably, the dosage form of the preparation is an emulsion, tablet, capsule, oral liquid, granule, powder, syrup or injection.

[0073] Preferably, the injectable preparation is an intravenous injection preparation, an intramuscular injection preparation, a subcutaneous injection preparation, an intradermal injection preparation, or an intraperitoneal injection preparation.

[0074] The fifth aspect of the present invention provides a method for preventing and / or treating platelet-related diseases, the method comprising administering to a desired subject hydroxysaffron yellow A and / or anagrelide or a pharmaceutically acceptable salt thereof, a composition as described in the second aspect of the present invention, or a cassette as described in the third aspect of the present invention.

[0075] Preferably, the prevention and / or treatment of platelet-related diseases includes one or more selected from the group consisting of: (i) prevention and / or treatment of thrombocytosis; (ii) reduction of platelet count; (iii) prevention and / or treatment of diseases associated with elevated platelet count; (iv) inhibition of platelet activation; (v) prevention and / or treatment of diseases associated with platelet activation; (vi) inhibition of platelet aggregation and / or clustering; (vii) prevention and / or treatment of diseases associated with platelet aggregation and / or clustering; (viii) reduction of large platelet ratio; (ix) prevention and / or treatment of elevated large platelet ratio; and / or (x) prevention and / or treatment of diseases associated with elevated large platelet ratio.

[0076] Preferably, the object is a human or a non-human mammal.

[0077] Preferably, the non-human mammal is a mouse, rat, rabbit, cow, sheep, dog, or horse.

[0078] Preferably, the application includes oral administration or injection.

[0079] Preferably, the injection administration is intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection, or intraperitoneal injection.

[0080] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Attached Figure Description

[0081] Figure 1 The distance the rat blood traveled in the capillary for each treatment group is represented by the following values: Control refers to the blank control group; HSYA refers to the HSYA group; Adr refers to the Adr group; and Adr+HSYA refers to the Adr+HSYA group.

[0082] Figure 2 The distance the blood traveled in the capillary in each treatment group of rats is represented by the distance traveled. Control refers to the blank control group; HSYA refers to the HSYA group; Adr refers to the Adr group; and Adr+HSYA refers to the Adr+HSYA group. Compared with the blank control group, *P<0.05, **P<0.01; compared with the Adr group, # P<0.05, ## P<0.01.

[0083] Figure 3 The aggregation of platelet-rich plasma (PRP) in each treatment group is shown. Control refers to the blank control group; HSYA refers to the HSYA group; Adr refers to the Adr group; and Adr+HSYA refers to the Adr+HSYA group.

[0084] Figure 4 Giemsa staining was performed on platelet-rich plasma (PRP) for each treatment group, where Control refers to the blank control group; HSYA refers to the HSYA group; Adr refers to the Adr group; and Adr+HSYA refers to the Adr+HSYA group.

[0085] Figure 5The fluorescence intensity of the Fluo-3 AM calcium ion probe in platelets of each treatment group was detected by flow cytometry. Among them, Control refers to the blank control group; HSYA refers to the HSYA group; Adr refers to the Adr group; and Adr+HSYA refers to the Adr+HSYA group.

[0086] Figure 6 To use FlowJo V10 Figure 5 The fluorescence intensity of the intraplatelet calcium ion probe Fluo-3 AM in each treatment group was quantified. Control refers to the blank control group; HSYA refers to the HSYA group; Adr refers to the Adr group; and Adr+HSYA refers to the Adr+HSYA group. Compared with the blank control group, *P<0.05, **P<0.01; compared with the Adr group, # P<0.05, ## P<0.01. Detailed Implementation

[0087] This invention discovers that hydroxysaffron yellow A can reduce the number of platelets and the ratio of large platelets, as well as inhibit platelet aggregation and agglutination. It can be used to prevent and treat diseases related to elevated platelet counts, such as thrombocytosis, diseases related to elevated large platelet ratios, and diseases related to platelet activation, thereby preventing and treating platelet-related diseases.

[0088] the term

[0089] As used herein, the terms “comprising,” “including,” and “containing” are used interchangeably and include not only open-ended definitions but also semi-closed and closed definitions. The terms include “consisting of” and “substantially consisting of”.

[0090] In this invention, the term "prevention" means a method of preventing the onset of a disease and / or its accompanying symptoms or protecting a subject from acquiring a disease.

[0091] In this invention, the term "treatment" includes delaying and halting the progression of the disease, or eliminating the disease, and does not require 100% inhibition, eradication, and reversal. In some embodiments, hydroxysaffron yellow A reduces, inhibits, and / or reverses thrombocytosis by, for example, at least about 30%, at least about 50%, or at least about 80%, at least about 90%, or 100% compared to levels observed in the absence of hydroxysaffron yellow A.

[0092] use

[0093] This invention provides the use of hydroxysaffron yellow pigment A in the prevention and / or treatment of platelet-related diseases.

[0094] In a preferred embodiment of the present invention, the present invention provides the use of hydroxysaffron yellow pigment A for preparing a composition for one or more uses selected from the group consisting of: (i) prevention and / or treatment of thrombocytosis; (ii) reduction of platelet count; (iii) prevention and / or treatment of diseases associated with elevated platelet count; (iv) inhibition of platelet activation; (v) prevention and / or treatment of diseases associated with platelet activation; (vi) inhibition of platelet aggregation and / or agglomeration; (vii) prevention and / or treatment of diseases associated with platelet aggregation and / or agglomeration; (viii) reduction of large platelet ratio; (ix) prevention and / or treatment of elevated large platelet ratio; and / or (x) prevention and / or treatment of diseases associated with elevated large platelet ratio.

[0095] In a preferred embodiment of the invention, the thrombocytosis includes thrombocytosis induced by adrenaline administration.

[0096] In a preferred embodiment of the invention, the reduction in platelet count includes reducing the platelet count of the subject receiving adrenaline.

[0097] In a preferred embodiment of the invention, the increase in platelet count includes an increase in platelet count induced by adrenaline administration.

[0098] In a preferred embodiment of the invention, the platelet activation includes platelet activation induced by adrenaline administration.

[0099] In a preferred embodiment of the invention, the platelet aggregation and / or aggregation includes platelet aggregation and / or aggregation induced by adrenaline administration.

[0100] In a preferred embodiment of the invention, the reduction of large platelet ratio includes reducing the large platelet ratio of the subject receiving adrenaline.

[0101] In a preferred embodiment of the invention, the increased platelet-to-large platelet ratio includes an increased platelet-to-large platelet ratio induced by adrenaline administration.

[0102] Compositions, formulations and packaging

[0103] The present invention also provides a composition or formulation, preferably a pharmaceutical composition or formulation. The composition or formulation of the present invention further includes a pharmaceutically acceptable carrier.

[0104] As used herein, the term "pharmaceutically acceptable carrier" refers to one or more compatible solid, semi-solid, liquid, or gel fillers that are suitable for human or animal use and have sufficient purity and low toxicity.

[0105] It should be understood that the carrier described in this invention is not particularly limited and can be any material commonly used in the art, prepared by conventional methods, or purchased from the market. Pharmaceutically acceptable examples of carriers include cellulose and its derivatives (such as methylcellulose, ethylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, etc.), polyols (such as propylene glycol, glycerin, sorbitol, etc.), emulsifiers (such as Tween), wetting agents (such as sodium dodecyl sulfate), buffers, chelating agents, thickeners, pH adjusters, transdermal penetration enhancers, colorants, flavoring agents, stabilizers, antioxidants, preservatives, antibacterial agents, pyrogen-free water, etc.

[0106] The compositions or formulations described in this invention may also include other drugs, such as anagrelide or a pharmaceutically acceptable salt thereof.

[0107] Preferably, the composition or preparation of the present invention comprises hydroxysaffron yellow A and anagrelide or a pharmaceutically acceptable salt thereof.

[0108] Anagrelene or a pharmaceutically acceptable salt thereof, as described in this invention, is preferably anagrelene hydrochloride.

[0109] In a preferred embodiment of the invention, the weight ratio of the hydroxysaffron yellow A to the anagre or a pharmaceutically acceptable salt thereof is 0.1-40:1, more preferably 0.3-30:1, more preferably 0.5-20:1, more preferably 0.5-15:1, more preferably 1-10:1, more preferably 1-8:1, more preferably 1-5:1, more preferably 2-4:1, more preferably 2.5-3.5:1, and more preferably 3:1.

[0110] In this invention, the dosage form of the composition or formulation includes (but is not limited to) solid dosage forms, semi-solid dosage forms, or liquid dosage forms.

[0111] In this invention, the dosage form of the composition or preparation includes (but is not limited to) oral preparations, injectable preparations, or topical preparations.

[0112] Typically, the injectable formulation is an intravenous injection formulation, an intramuscular injection formulation, a subcutaneous injection formulation, an intradermal injection formulation, or an intraperitoneal injection formulation.

[0113] Pharmaceutical formulations should be matched with the route of administration, which involves administering a therapeutically effective amount of the drug to the intended recipient (e.g., a human or non-human mammal). As used herein, the term "therapeutically effective amount" refers to an amount that is functionally or activityily produced in humans and / or animals and is acceptable to them. Those skilled in the art will understand that the "therapeutically effective amount" can vary depending on the form of the pharmaceutical formulation, the route of administration, the excipients used, the severity of the disease, and whether it is used in combination with other drugs, all of which are within the scope of the skill of a skilled physician.

[0114] The main technical effects achieved by this invention include:

[0115] This invention discovers that hydroxysaffron yellow A can reduce the number of platelets and the ratio of large platelets, as well as inhibit platelet aggregation and agglutination. It can be used to prevent and treat diseases related to elevated platelet counts, such as thrombocytosis, diseases related to elevated large platelet ratios, and diseases related to platelet activation, thereby preventing and treating platelet-related diseases.

[0116] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that the following specific embodiments are based on the present technical solution and provide detailed implementation methods and specific operation processes, but the scope of protection of the present invention is not limited to these embodiments.

[0117] Example 1

[0118] 1. Reagents

[0119] Hydroxysafflor yellow A was purchased commercially; its English name is Hydroxysafflor yellow A, abbreviated as HSYA, and its CAS number is 78281-02-4.

[0120] Adrenaline is commercially available and is abbreviated as Adr. Its CAS number is 51-43-4.

[0121] Anagrelide Hydrochloride, abbreviated as Ana, was purchased commercially and has the CAS number 58579-51-4.

[0122] 2. Laboratory animals

[0123] Male SPF-grade Sprague-Dawley (SD) rats, weighing 250–300 g, were housed in an isolated environment with a 12–12 h diurnal cycle, with the room temperature maintained at (24±1) ℃. They had free access to water and food and were allowed to acclimatize to the environment for one week before the experiment was conducted.

[0124] 3. Effect Experiment

[0125] 3.1 Hydroxysafflower yellow A (HSYA) reduces platelet count and the ratio of large platelets, and inhibits platelet aggregation.

[0126] Forty male SPF-grade SD rats were randomly divided into four groups (n=10): a blank control group, an HSYA group, an Adr group, and an Adr+HSYA group. Rats in the HSYA and Adr+HSYA groups were intraperitoneally injected with a saline dispersion of hydroxysaffron yellow A (3 mg / kg bw), while rats in the blank control and Adr groups were intraperitoneally injected with the same volume of saline. Administration was once daily for 5 consecutive days. Afterward, the rats in each group underwent the following treatment: rats in the Adr and Adr+HSYA groups were subcutaneously injected with a saline dispersion of epinephrine (1 mg / kg bw), while rats in the blank control and HSYA groups were subcutaneously injected with the same volume of saline. One hour later, blood was collected from each group for complete blood count, blood coagulation rate, and platelet aggregation rate (PAG) testing. The testing methods are as follows:

[0127] Complete blood count (CBC) test:

[0128] One mL of blood was collected from each group of rats for routine blood tests. Platelet count (PLT), mean platelet volume (MPV), platelet distribution width (PDW), and large platelet ratio (P-LCR) were measured using a blood analyzer. The results of routine blood tests for each group of rats are shown in Table 1.

[0129] Blood coagulation rate detection:

[0130] 100 μl of blood from each group of rats was placed in a 200 μL PCR tube. A capillary glass rod (0.1 mm in diameter) was vertically inserted into the tube at 37°C, and the height of the blood rise in the capillary within 1 minute was measured. The results are as follows: Figure 1 and Figure 2 As shown.

[0131] Platelet aggregation rate (PAG) test:

[0132] Rat blood was collected, heparin sodium was added for anticoagulation, and the mixture was centrifuged at 100g at room temperature for 10 minutes. The supernatant was collected to obtain platelet-rich plasma (PRP). The aggregation of platelet-rich plasma (PRP) in each treatment group is shown in the figure. Figure 3As shown. Giemsa staining was performed on platelet-rich plasma (PRP). The staining procedure was as follows: PRP from each treatment group was collected, and 50 μL of PRP was added to a clean glass slide to prepare a smear. After the PRP dried on the slide, 80 μL of Giemsa staining solution was added, and staining was performed for 30 seconds. Then, 160 μL of PBS was added, and staining continued for 5 minutes. After staining, the slides were rinsed with water for 3 minutes. After drying, the slides were observed using an optical microscope. The Giemsa staining of platelet-rich plasma (PRP) is shown below. Figure 4 As shown.

[0133] The intracellular calcium content in platelet cytoplasm was measured using the calcium ion probe Fluo-3 AM via flow cytometry. 2+ Concentration. Fluorescence intensity of the intraplatelet calcium ion probe Fluo-3 AM in each treatment group, as determined by flow cytometry, is shown below. Figure 5 As shown, the results of quantifying the fluorescence intensity of the intracellular calcium ion probe Fluo-3 AM using FlowJo V10 are as follows: Figure 6 As shown.

[0134] Table 1. Blood routine tests of rats in each treatment group

[0135] project Blank control group HSYA Group Adr Group Adr+HSYA group <![CDATA[PLT(×10 9 / L)]]> 510±46 <![CDATA[326±42 ## ]]> 536±73 <![CDATA[371±67 ## ]]> MPV(fl) 7.5±0.2 7.3±0.1 7.5±0.1 7.2±0.2 PDW (%) 8.2±0.2 7.8±0.2 8.2±0.1 7.8±0.2 P-LCR (%) 6.2±0.2 <![CDATA[5.3±0.3 # ]]> <![CDATA[7.9±0.1 * ]]> <![CDATA[7.0±0.2 # ]]> PAG (%) 17.6±5.9 8.9±4.4 62.9±14.5 34.6±9.7

[0136] Notes: PLT refers to blood platelet count; MPV refers to mean platelet volume; PDW refers to platelet distribution width; P-LCR refers to platelet-larger cell ratio; PAG refers to platelet aggregation rate. *P<0.05 and **P<0.01 indicate statistically significant differences compared to the blank control group. # P<0.05 and ## P<0.01 means that the difference is statistically significant compared with the Adr group.

[0137] As can be seen from the blood routine test results in Table 1, hydroxysaffron yellow pigment A (HSYA) can significantly reduce the number of platelets and the ratio of large platelets in peripheral blood. Therefore, hydroxysaffron yellow pigment A (HSYA) has excellent therapeutic effects on thrombocytosis and increased large platelet ratio, such as thrombocytosis and increased large platelet ratio caused by adrenaline (Adr) administration.

[0138] Table 1 shows the platelet aggregation rate (PAG) experimental data, indicating that hydroxysaffron yellow A (HSYA) effectively inhibits platelet aggregation. The distance the blood travels in the capillaries of rats in each treatment group is shown in the table. Figure 1 and Figure 2 As shown, Figure 1 and Figure 2 Treatment with hydroxysaffron yellow A (HSYA) significantly upregulated the rise of blood within capillaries, indicating that HSYA can significantly reduce blood aggregation rate. Figure 3 and Figure 4 The platelet aggregation data from the different treatment groups showed that hydroxysaffron yellow A (HSYA) significantly reduced platelet aggregation. Therefore, hydroxysaffron yellow A (HSYA) can effectively inhibit platelet aggregation and thus can be used to inhibit platelet activation.

[0139] The results of flow cytometry analysis of platelet calcium ion content in each treatment group are as follows: Figure 5 and Figure 6 As shown. Figure 5 and Figure 6 The results showed that HSYA could significantly reduce the content of intraplatelet calcium ions and significantly inhibit adrenaline-induced upregulation of intraplatelet calcium ions.

[0140] 3.2-Hydroxysafflower yellow A (HSYA) combined with anagrelide hydrochloride (Ana) reduces platelet count.

[0141] Fifty male SPF-grade SD rats were randomly divided into 5 groups (n=10): blank control group, Adr group, Adr+HSYA group, Adr+Ana group, and Adr+HSYA+Ana group. Rats in the Adr+HSYA group were intraperitoneally injected with hydroxysaffron yellow A saline dispersion (hydroxysaffron yellow A dosage: 3.0 mg / kg bw); rats in the Adr+Ana group were intraperitoneally injected with anagrelide hydrochloride (anagrelide hydrochloride dosage: 1.0 mg / kg bw); rats in the Adr+HSYA+Ana group were intraperitoneally injected with hydroxysaffron yellow A and anagrelide hydrochloride saline dispersion (hydroxysaffron yellow A dosage: 1.5 mg / kg bw, anagrelide hydrochloride dosage: 0.5 mg / kg bw); and rats in the blank control group and Adr group were injected with anagrelide hydrochloride saline dispersion (hydroxysaffron yellow A dosage: 1.5 mg / kg bw, anagrelide hydrochloride dosage: 0.5 mg / kg bw). Each rat was intraperitoneally injected with the same volume of physiological saline once a day for 5 consecutive days. After that, the rats in each group were treated as follows: each rat in the Adr group, Adr+HSYA group, Adr+Ana group and Adr+HSYA+Ana group was subcutaneously injected with adrenaline saline dispersion (the dosage of adrenaline was 1 mg / kg bw), and each rat in the blank control group was subcutaneously injected with the same volume of physiological saline. One hour later, blood was collected from each group of rats, and the platelet count (PLT) in the blood of each group of rats was measured by a blood analyzer. The platelet count (PLT) test results of each treatment group of rats are shown in Table 2.

[0142] Table 2 Platelet counts (PLT) in rats of each treatment group

[0143] Processing group <![CDATA[PLT(×10 9 / L)]]> Blank control group 510±46 Adr Group 536±73 Adr+HSYA group <![CDATA[371±67 ## ]]> Adr+Ana group <![CDATA[428±43 ## ]]> Adr+HSYA+Ana group <![CDATA[342±31 ## ]]>

[0144] Note: PLT refers to blood platelet count; # P<0.05 and ## P<0.01 means that the difference is statistically significant compared with the Adr group.

[0145] As shown in Table 2, the platelet counts (PLT) of rats in each group indicate that hydroxysaffron yellow A (HSYA) and anagrelide hydrochloride can reduce the number of platelets in peripheral blood. Compared with hydroxysaffron yellow A (HSYA) or anagrelide hydrochloride alone, the combination of hydroxysaffron yellow A and anagrelide hydrochloride has a superior effect in reducing platelet count, and exhibits a synergistic effect in reducing platelet count. Therefore, hydroxysaffron yellow A and anagrelide hydrochloride can synergistically provide excellent therapeutic effects for thrombocytosis.

[0146] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. The use of a hydroxysaffron yellow pigment A, characterized in that, Used to prepare a composition for the prevention and / or treatment of thrombocytosis; The thrombocytosis mentioned is thrombocytosis induced by adrenaline administration; The prevention and / or treatment targets are subjects who have already received adrenaline, and the platelet count of the subjects who received adrenaline is elevated.

2. The use as described in claim 1, characterized in that, The composition is a pharmaceutical composition.

3. The use as described in claim 1, characterized in that, The composition also includes a pharmaceutically acceptable carrier.

4. The use as described in claim 1, characterized in that, The dosage form of the composition is an oral preparation, a topical preparation, or an injectable preparation.

5. The use as described in claim 1, characterized in that, The objects described are human or non-human mammals.

Images